Fig. 4

EZH2 is required for YAP-mediated repression of JMJD3. A Relative protein expression of EZH2, JMJD3 and H3K27me3 after transfection with siEZH2-1,2, or negative control in two cell lines. B Relative protein expression of YAP, EZH2, JMJD3 and H3K27me3 in the oeNC, oeYAP, and oeYAP+ siEZH2 groups across two cell lines. C Co-IP experiment in HT29 cells was performed to validate the combination between YAP and EZH2. D Immunofluorescence colocalization analysis of Caco-2 cells stained with anti-EZH2 (green) and anti-YAP (red) antibodies; nuclei were counterstained with DAPI (blue). Quantification of immunofluorescence data shown on right. E, F ChIP-PCR analysis to validate the enrichment of H3K27me3 on the JMJD3 promoter in E HT29 cells and F Caco-2 cells transfected with siEZH2, siYAP or siNC, with IgG used as a negative control